1. Field of the Invention
The present invention relates to the use of medicamentous vaccine preparations with a therapeutic or prophylactic use intended to treat or prevent, in malignant tumors, immune disorders in particular immunosuppression and apoptosis of the immune or vascular cells such as angiogenesis, induced by extracellular factors, cytokines or other regulation factors in particular transcriptional factors, abnormally produced by the cancer cells or the stromal cells.
2. Description of the Related Art
Conventional treatments of cancers whether they are of viral origin, induced by retroviruses, or EBV or HPV or also the hepatitis viruses, or of chronic origin, due to asbestos or to benzene derivatives, whether they are of epithelial (carcinomas) or conjunctive (sarcomas) type or also of the blood (lymphomas) comprise the surgical removal of tumors usually combined with chemotherapy and/or radiotherapy.
Although effective for certain cancers, particularly when carried out in the early stages, these often poorly tolerated treatments are inadequate and relapses and metastases compromise the progress of patients.
This is why when scientists in the 80's and 90's cloned and purified tumor-associated antigens (TAA) or tumor-specific antigens (TSA) in cancer cells originating from numerous malignant tumors (cancer of the breast, prostate, colon and rectum, cervix, ATL lymphoma), numerous experiments and clinical trials of anti-cancer vaccination (Dvorak E. Experimental design for vaccine preparations against human malignant tumors. Med Hypotheses (1986) 20:429-52, Houghton A N. On course for a cancer vaccine. Lancet (1995) 345:1384-5, Herlyn D, Linnenbach A, Koprowski H, Herlyn M. Epitope- and antigen-specific cancer vaccines. Int Rev Immunol (1991) 7:245-57, antigen-specific cancer vaccines. Int Rev Immunol (1991) 7:245-57, Ostankovitch M, Choppin J, Guillet J G. Tumor cell antigenicity: cancers and vaccines. Rev Prat (1995) 45:1921-6, Zhu M Z, Marshall J, Cole D, Schlom J, Tsang K Y. Specific cytolytic T-cell responses to human CEA from patients immunized with recombinant avipox-CEA vaccine. Clin Cancer Res (2000) 6:24-33, Tsunoda T, Tanimura H, Yamaue H, Tanaka H, Matsuda K. Tumor specific CTL therapy for advanced cancer and development for cancer vaccine. Hepatogastroenterology (1999) 46:1287-92), using TAA and TSA as antigens were carried out, aiming to specifically destroy the malignant cells which carry these antigens thanks to the action of killer cells, particularly cytolytic lymphocytes (CTL), carriers of specific receptors, induced by the vaccinal immune reaction.
Clinical trials using such vaccines carried out in patients carrying different tumors (melanoma, cancer of the breast, colorectal cancer, cancer of the bladder etc.) have made it possible to establish the following facts:                The anti-cancer vaccine preparations containing the tumorous antigens (TAA or TSA) presented in different forms have been well tolerated and have generally not caused regional or systemic complications.        Such vaccine preparations can induce in patients an immune reaction of CTL type (Tsunoda T, Tanimura H, Yamaue H, Tanaka H, Matsuda K. Tumor specific CTL therapy for advanced cancer and development for cancer vaccine. Hepatogastroenterology (1999) 1:1287-92, Schwaab T, Heaney J A, Schned A R, Harris R D, Cole B F, Noelle R J, Phillips D M, Stempkowski L, Ernstoff M S. A randomized phase II trial comparing two different sequence combinations of autologous vaccine and human recombinant interferon gamma and human recombinant interferon alpha2B therapy in patients with metastatic renal cell carcinoma: clinical outcome and analysis of immunological parameters. J Urol (2000) 163:1322-7, Steller M A, Gurski K J, Murakami M, Daniel R W, Shah K V, Celis E, Sette A, Trimble E L, Park R C, Marincola F M. Cell-mediated immunological responses in cervical and vaginal cancer patients immunized with a lipidated epitope of human papillomavirus type 16 E7. Clin Cancer Res (1998) 4:2103-9, capable in vitro of specifically destroying the cellular targets which carry TAA or TSA epitopes complexed with the Major Histocompatibity Complex.        On the other hand, to date, no phase III clinical trial has been able to show that these vaccine preparations, aiming to specifically destroy the cancerous cells by the differentiation of killer cells, were effective.        
Therefore, since 1992, after Levine (The p53 tumor suppressor gene and gene product. Princess Takamatsu Symp (1989) 20:221-30) as well as other scientific teams had showed that the native p53 protein which has reparative effects on the DNA strands and immunosuppressive effects on the cell cycle or a mutant of this protein was abundantly produced and accumulated in malignant tumors, the same Levine proposes carrying out a vaccination using the p53 protein, appearing to be a tumor-associated antigen (TAA). This was presented at the surface of dendritic cells (DC) or added into a bacterial vector (BCG type) in such a way as to induce an immune reaction of CTL type directed against the cancerous cells (See also WO-A-94/02167).
In support of this patent application, scientific publications show the beneficial role of killer cells and the pejorative role of specific antibodies in the development of malignant tumors (Theobald M, Biggs J, Dittmer D, Levine A J, Sherman L A. Targeting p53 as a general tumor antigen. Proc Natl Acad Sci USA (1995) 92:11993-7, Roth J. et al, p53 as a target for cancer vaccines: recombinant canarypox virus vectors expressing p53 protect mice against lethal tumor cell challenge, 1996, Proc Natl Acad Sci USA.; 93:4781-6).
Following Levine, other teams modifying the vector or the adjuvant of the p53 immunogen have filed ten or so patent applications on the use of new galenic formulations of anti-p53 vaccine also aiming to induce the formation of CTL killer T cells targeting the cancer cells expressing the p53 protein.
The experimental trials associated with these anti-p53 vaccines have shown the innocuousness and the immunogenicity assessed by the appearance of anti-p53 killer cells. In addition, the only clinical trial of anti-p53 vaccination which has been carried out and published confirmed the innocuousness and the immunogenicity of the vaccine. However no phase III trial has been able to validate the effectiveness of this vaccine strategy.
The Applicant surprisingly discovered that the immunosuppression and the angiogenesis of the microenvironment of cells infected by some viruses such as HIV-1 and the microenvironment of cancerous cells provide a rational explanation for the lack of effectiveness of these vaccine strategies, as these previous strategies target the cancerous cell and not the disturbance of its microenvironment.
Now, to date the treatments have all aimed to directly kill the cancer T cells themselves, this is to say the parenchymetous cells, the Applicant has found that it was just as judicious or even more judicious to combat the molecules produced in the extracellular (stromal) microenvironment of the tumor and encouraging the development of the latter.
It should be remembered that any tissue or tumor is formed from parenchymetous cells which bathe in a microenvironment called stroma. This stroma is itself constituted by stromal cells (which can be immune, endothelial, or fibroblastic cells) and an extracellular medium.
The works of the Applicant have shown in fact that soluble factors secreted by the cells infected by HIV-1, in particular the Tat protein or by the immune cells of patients infected by HIV in particular IFNα and TGFβ or produced by cancer cells, such as the E7 protein of HPV in cancer of the cervix or the Tax protein of HTLV1 in the ATL leukaemias or the p53 protein in colorectal cancer, had immunosuppressive properties capable of inhibiting the cellular immune reactions in tumors and because of this explained the ineffectiveness of previous vaccines.
Bibliographical study has made it possible to confirm these observations by the Applicant, confirming the presence of immunosuppressive factors released into the extracellular medium of malignant tumors:
Some of these as yet unidentified factors were produced by                colorectal cancer cells (Ebert E C, Roberts Al, O'Connell S M, Robertson F M, Nagase H. Characterization of an immunosuppressive factor derived from colon cancer cells. J Immunol. (1987) 138:2161-8 or Remacle-Bonnet M M, Pommier F J, Kaplanski S, Rance R J, Depieds R C. Inhibition of normal allogenic lymphocyte mitogenesis by a soluble inhibitor extracted from human colonic carcinoma. J Immunol (1976) 117:1145-51,        glioblastoma cells (29-Fontana A, Hengartner H, de Tribolet N, Weber E. Glioblastoma cells release interleukin 1 and factors inhibiting interleukin 2-mediated effects. J Immunol. (1984) 132:183744),        melanomas (30.Hersey P, Bindon C, Czerniecki M, Spurling A, Wass J, McCarthy W H. Inhibition of interleukin 2 production by factors released from tumor cells. J Immunol. (1983) 131:2837-42), or malignant ascites (Tamura K, Shibata Y, Matsuda Y, Ishida N. Isolation and characterization of an immunosuppressive acidic protein from ascitic fluids of cancer patients. Cancer Res. (1981) 41:3244-52, Oh S K, Moolten F L. Non specific immunosuppressive factors in malignant ascites: further characterization and possible relationship to erythrocyte receptors of human peripheral T cells. J Immunol. (1981) 127:2300-7).        
Other transcriptional regulation factors, as reported above, are of cellular origin such as the p53 protein, accumulated in some malignant tumors, in particular colorectal tumors (Remvikos Y, Tominaga O, Hammel P, Laurent-Puig P, Salmon R J, Dutrillaux B, Thomas G. Increased p53 protein content of colorectal tumors correlates with poor survival. Br J Cancer 1992 66:758-64, Gan H, Ouyang Q, Wang Y. Expression of p53 protein in colorectal cancer and its relationship to cell proliferative activity and prognosis. Chung Hua Chung Liu Tsa Chih (1996) 18:244-6). The p53 protein, released by active transport by the secretion pathways not using the peptide signal or by passive diffusion is present in the extracellular medium, and it has been isolated by chromatography on glass fibre from serum from cancer patients (Zusman I, Sandier B, Gurevich P, Zusman R, Smirnoff P, Tendler Y, Bass D, Shani A, Idelevich E, Pfefferman R, Davidovich B, Huszar M, Glick J. Comparative study of the role of serum levels of p53 antigen and its tumor cell concentration in colon cancer detection. Hum Antibodies Hybridomas. (1996):123-8, Sandler B, Smirnoff P, Tendelr Y, Zinder O, Zusman R, Zusman I. Specificity of polyclonal anti-p53 IgG for isolation of the soluble p53 antigen from human serum. Int J Mol. Med. 1998 1:767-70).